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Circ_072697 knockdown promotes advanced glycation end products-induced cell proliferation and migration in HaCaT
Ming Tian1, Jiajun Tang2, Rong Huang3,4
1Department of Burn, Wound Healing Center, Shanghai Burn Institute, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China.
Objective:
Diabetes foot ulcer (DFU) is a serious complication of diabetes, which can lead to significant mortality and amputation rate. Our previous study found circ_072697 was highly expressed in DFU tissues, but the regulatory mechanism of circ_072697 in DFU remains unclear.
Methods:
The relative expressions of circ_072697, miR-3150a-3p, and KDM2A in DFU patients or advanced glycation end products (AGEs)-treated HaCaT cells (used as DFU cell model) were determined by using qRT-PCR. Cell proliferation and migration abilities were determined by using CCK-8 and Transwell assays. The interaction between miR-3150a-3p with circ_072697 or KDM2A were verified by RNA immunoprecipitation (RIP) and dual-luciferase reporter assays. Furthermore, the protein expression of genes involved in MAPK signaling pathway was detected by western blot.
Results:
The expression of circ_072697 was significantly upregulated in DFU tissues, while the expression of miR-3150a-3p was downregulated. Circ_072697 knockdown promoted the proliferation and migration of AGEs-treated HaCaT cells. miR-3150a-3p was confirmed as a target of circ_072697 and its inhibitor reversed the promotion effects of circ_072697 knockdown on biological behavior of cells. In addition, KDM2A was considered as a target of miR-3150a-3p and it was highly expressed in DFU samples. Importantly, circ_072697 could regulate KDM2A expression through sponging miR-3150a-3p, and this axis had effect on the MAPK signaling pathway.
Conclusions:
Overall, circ_072697 regulated the biological behaviors of keratinocytes in DFU via miR-3150a-3p/KDM2A axis and MAPK signaling pathway, revealing a new insight into the pathogenesis and potential therapeutic targets of DFU.
Insights
Circular RNA circ_072697 promotes diabetes foot ulcer (DFU) progression by sponging miR-3150a-3p, upregulating KDM2A, and affecting the MAPK pathway. This highlights circ_072697 as a potential therapeutic target for DFU.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Diabetes foot ulcer (DFU) presents a significant health challenge, associated with high mortality and amputation rates.
- Previous research identified elevated circ_072697 expression in DFU tissues, but its precise role and regulatory mechanisms remain largely unknown.
Purpose of the Study:
- To elucidate the regulatory mechanism of circ_072697 in the pathogenesis of diabetes foot ulcers.
- To investigate the interaction between circ_072697, miR-3150a-3p, and KDM2A in DFU development.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) to assess gene expression.
- Cell proliferation and migration assays (CCK-8, Transwell).
- RNA immunoprecipitation (RIP) and dual-luciferase reporter assays to confirm molecular interactions.
- Western blot analysis to detect protein expression in the MAPK signaling pathway.
Main Results:
- Circ_072697 was significantly upregulated, while miR-3150a-3p was downregulated in DFU tissues and AGEs-treated HaCaT cells.
- Knockdown of circ_072697 inhibited cell proliferation and migration; miR-3150a-3p inhibition reversed these effects.
- KDM2A was identified as a target of miR-3150a-3p and was highly expressed in DFU samples.
- Circ_072697 regulates KDM2A via sponging miR-3150a-3p, impacting the MAPK signaling pathway.
Conclusions:
- Circ_072697 promotes DFU progression by modulating the miR-3150a-3p/KDM2A axis and influencing the MAPK signaling pathway.
- This study reveals a novel molecular mechanism underlying DFU pathogenesis.
- The circ_072697/miR-3150a-3p/KDM2A axis represents a potential therapeutic target for DFU treatment.
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